Zavil'gel'skiĭ G B, Mershavka V Iu, Iusifov T N, Belogurov A A
Mol Biol (Mosk). 1984 Nov-Dec;18(6):1590-6.
The host-controlled K-restriction of unmodified phage lambda is ten to hundred-fold alleviated in the E. coli K12 strain, carring plasmid pKM101 of N-incompatibility group. By restriction mapping Tn5 insertion in pKM101, which reduced pKM101-mediated alleviation of K-restriction, was shown to by located within BglII-B-fragment approximately 9 kb anticlockwise from the EcoRI-site of pKM101. We have termed the gene(s) promoting the alleviation of K-restriction ARD (Alleviation of Restriction of DNA). It was shown that (i) plasmid pKM101-mediated alleviation of K-restriction did not depend on bacterial genes LexA, RecBC, umuC and plasmid gene muc; (ii) ard gene did not mediate EcoK type modification of DNA and did not enhance the modification activity of EcoK system in a way similar to that observed with RAL gene of phage lambda. Action of Ard gene of plasmid pKM101 is highly specific: alleviation of restriction of DNA lambda takes place only in K-strains of E. coli and is practically absent in B-strains and also in E. coli strains which have restricting enzymes of 11 type, EcoRI and EcoRIII.
在携带N不相容群质粒pKM101的大肠杆菌K12菌株中,未修饰的噬菌体λ的宿主控制的K限制被减轻了10到100倍。通过对pKM101中Tn5插入进行限制酶切图谱分析,发现降低pKM101介导的K限制减轻作用的Tn5插入位于pKM101 EcoRI位点逆时针方向约9 kb处的BglII - B片段内。我们将促进K限制减轻的基因命名为ARD(DNA限制减轻)。结果表明:(i)质粒pKM101介导的K限制减轻不依赖于细菌基因LexA、RecBC、umuC和质粒基因muc;(ii)ard基因不介导DNA的EcoK型修饰,也不像噬菌体λ的RAL基因那样以类似方式增强EcoK系统的修饰活性。质粒pKM101的Ard基因作用具有高度特异性:DNAλ的限制减轻仅在大肠杆菌的K菌株中发生,而在B菌株以及具有11型限制酶EcoRI和EcoRIII的大肠杆菌菌株中几乎不存在。